In Mediterranean countries, forage grasses and legumes are commonly grown in mixture because of their ability to increase herbage yield and quality compared with monocrop systems. However, the benefits of intercropping over a monocrop system are not always realized because the efficiency of a grass–legume mixture is strongly affected by agronomic factors. The present study evaluated productivity, N2 fixation, N transfer, and N recovery of berseem clover (Trifolium alexandrinum) grown in pure stand and in mixture with annual ryegrass (Lolium multiflorum) under high or low defoliation frequencies and varying plant arrangements (sowing in the same row or in alternating rows). On average, the berseem–ryegrass mixtures resulted in a greater yield and N yield than the monocrops. When mixed together, ryegrass was more efficient than berseem at absorbing soil N, increasing the reliance of berseem on N2 fixation. Both defoliation management and plant arrangement affected forage yield and the quality of the mixture, modifying the proportion of the two components, the N content of the forage, and the symbiotic N2 fixation of the legume. Reducing the proximity between plants of the two species may benefit the weaker component of the mixture. No apparent transfer of fixed N from berseem to ryegrass was detected